Premature infants admitted to the neonatal intensive care unit (NICU) require up to several hundred procedures during their hospitalization. Many of these are tissue-damaging procedures (TDPs) known to cause pain. Through funding from NINR, we found that TDPs not only caused pain but also increased markers of ATP degradation and oxidative stress. Based on this finding, we examined the effect of oral sucrose, to determine if this commonly used analgesic reduces biochemical markers of ATP degradation and oxidative stress. We hypothesized that since oral sucrose was documented to significantly reduce pain scores, then administration of this analgesic will also decrease markers of ATP degradation and oxidative stress. However, we observed the opposite effect. Although a single dose of oral sucrose reduced behavioral markers of pain, it significantly increased biochemical markers of ATP degradation (hypoxanthine, uric acid) and oxidative stress (allantoin) over time. More importantly, the effect of oral sucrose on breakdown markers of ATP were enhanced and were significantly higher in neonates that were intubated or were receiving more than 30% FiO2 . These findings lead to the question: If oral sucrose is ineffective in reducing the biochemical effects of procedural pain, what intervention or groups of intervention will decrease both behavioral markers of procedural pain and reduce ATP utilization and oxidative stress in premature neonates? For this RO1 renewal, we will answer this question by examining the individual and additive effects of two commonly used interventions: (a) 30% oral glucose (b) facilitated tucking c) 30% oral glucose and facilitated tucking. These interventions are commonly used but their effects on cellular bioenergetics and oxidative stress is unknown. Findings from this study will provide clinicians with evidence-based interventions that will both decrease behavioral signs of pain and biochemical markers of ATP utilization and oxidative stress. This will lead the way toward future clinical trials that will examine the relationship between prevention of pain and organ injury in the most fragile members of our society.